Heparan sulfate (HS) is an essential glycan that is present in large quantities on the cell surface and in the extracellular matrix. HS participates ina variety of physiological and pathophysiological functions, including blood coagulation, inflammatory response and embrynic development. HS is a highly sulfated polysaccharide. Heparin, a special form of HS, is a commonly used anticoagulant drug. The uniquely distributed sulfation pattern of HS is believed to regulate its functional specificity. The wide range of biological functions of HS attracts considerable interest in understanding the mechanism for controlling the biosynthesis of HS. HS biosynthesis involves a series of specialized HS sulfotransferases, glycosyltransferases and an epimerase. Using HS biosynthetic enzymes, our labs can produce an array of HS with unique sulfation patterns and functions. Our success has proved the feasibility for studying the control of the biosynthesis of specific sulfated saccharide sequences. Our hypothesis is that the HS biosynthesis is controlled by a series of up- stream modification steps involving N-sulfation and C5-epimerization with or without concurrent 2-O-sulfation. In this proposal, we will investigate the mechanism used by C5-epimerase and 2-O-sulfotransferase to control the synthesis of specific saccharide sequences that exhibit the anticoagulant activity and the activity in stimulating growth factors and growth factor receptor mediated cell proliferation. We will use structurally defined oligosaccharide substrates with precisely controlled N-sulfation patterns to prove the substrate specificity of C5-epimerase and 2-O-sulfotransferase. Finally, we plan to prove a unique biosynthetic pathway for preparing a highly sulfated domain that is commonly found in heparin. The success of this project will lead a comprehensive new approach to investigate the biosynthesis of HS, potentially leading to the development of novel HS-based therapeutics.

Public Health Relevance

Heparan sulfate (HS) is is a highly sulfated polysaccharide. HS displays a wide range of biological activities, including anticoagulant, antiinflammation and controlling the embryonic development. Currently, the biosynthetic mechanism of heparan sulfate is unknown. In this proposal, we plan to investigate the biosynthesis of heparan sulfate using structurally defined oligosaccharide substrates. The success of this project could provide a method to develop new heparan sulfate-based therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01GM102137-03
Application #
8853290
Study Section
Intercellular Interactions Study Section (ICI)
Program Officer
Marino, Pamela
Project Start
2013-09-01
Project End
2016-05-31
Budget Start
2015-06-01
Budget End
2016-05-31
Support Year
3
Fiscal Year
2015
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Type
Schools of Pharmacy
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
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